Osmoprotective effects of supplemental epidermal growth factor in an ex vivo multilayered human conjunctival model under hyperosmotic stress
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- 作者：Jae-hyung Kim (1)
Soon-Suk Kang (2)
Eun Soon Kim (2)
Jae Yong Kim (2)
Myoung Joon Kim (2)
Hungwon Tchah (2) - 关键词：Dry eye ; Epidermal growth factor ; Multilayered culture ; Osmoprotective effect
- 刊名：Graefe's Archive for Clinical and Experimental Ophthalmology
- 出版年：2013
- 出版时间：August 2013
- 年：2013
- 卷：251
- 期：8
- 页码：1945-1953
- 全文大小：370KB
- 参考文献：1. Brewitt H, Sistani F (2001) Dry eye disease: the scale of the problem. Surv Ophthalmol 45(Suppl 2):S199-02 CrossRef
2. Foulks GN (2008) Pharmacological management of dry eye in the elderly patient. Drugs Aging 25:105-18 CrossRef
3. Behrens A, Doyle JJ, Stern L, Chuck RS, McDonnell PJ, Azar DT, Dua HS, Hom M, Karpecki PM, Laibson PR, Lemp MA, Meisler DM, Del Castillo JM, O’Brien TP, Pflugfelder SC, Rolando M, Schein OD, Seitz B, Tseng SC, van Setten G, Wilson SE, Yiu SC (2006) Dysfunctional tear syndrome: a Delphi approach to treatment recommendations. Cornea 25:900-07 CrossRef
4. WorkShop DE (2007) The definition and classification of dry eye disease: report of the Definition and Classification Subcommittee of the International Dry Eye WorkShop. Ocul Surf 5:75-2 CrossRef
5. Lemp MA (2008) Management of dry eye disease. Am J Manag Care 14:S88-01
6. Pflugfelder SC (2004) Antiinflammatory therapy for dry eye. Am J Ophthalmol 137:337-42 CrossRef
7. WorkShop DE (2007) Management and therapy of dry eye disease: report of the Management and Therapy Subcommittee of the International Dry Eye WorkShop. Ocul Surf 5:163-78 CrossRef
8. Latkany R (2008) Dry eyes: etiology and management. Curr Opin Ophthalmol 19:287-91 CrossRef
9. Narayanan S, Manning J, Proske R, McDermott AM (2006) Effect of hyperosmolality on beta-defensin gene expression by human corneal epithelial cells. Cornea 25:1063-068 CrossRef
10. Bildin VN, Yang H, Crook RB, Fischbarg J, Reinach PS (2000) Adaptation by corneal epithelial cells to chronic hypertonic stress depends on upregulation of Na:K:2Cl cotransporter gene and protein expression and ion transport activity. J Membr Biol 177:41-0 CrossRef
11. Gilbard JP, Carter JB, Sang DN, Refojo MF, Hanninen LA, Kenyon KR (1984) Morphologic effect of hyperosmolarity on rabbit corneal epithelium. Ophthalmology 91:1205-212
12. Li DQ, Chen Z, Song XJ, Luo L, Pflugfelder SC (2004) Stimulation of matrix metalloproteinases by hyperosmolarity via a JNK pathway in human corneal epithelial cells. Invest Ophthalmol Vis Sci 45:4302-311 CrossRef
13. Li DQ, Luo L, Chen Z, Kim HS, Song XJ, Pflugfelder SC (2006) JNK and ERK MAP kinases mediate induction of IL-1beta, TNF-alpha and IL-8 following hyperosmolar stress in human limbal epithelial cells. Exp Eye Res 82:588-96 CrossRef
14. Pan Z, Wang Z, Yang H, Zhang F, Reinach PS (2011) TRPV1 activation is required for hypertonicity-stimulated inflammatory cytokine release in human corneal epithelial cells. Invest Ophthalmol Vis Sci 52:485-93 CrossRef
15. Enriquez-de-Salamanca A, Calder V, Gao J, Galatowicz G, García-Vázquez C, Fernández I, Stern ME, Diebold Y, Calonge M (2008) Cytokine responses by conjunctival epithelial cells: an in vitro model of ocular inflammation. Cytokine 44:160-67 CrossRef
16. Pflugfelder SC, Jones D, Ji Z, Afonso A, Monroy D (1999) Altered cytokine balance in the tear fluid and conjunctiva of patients with Sjogren’s syndrome keratoconjunctivitis sicca. Curr Eye Res 19:201-11 CrossRef
17. Shatos MA, Gu J, Hodges RR, Lashkari K, Dartt DA (2008) ERK/p44p42 mitogen-activated protein kinase mediates EGF-stimulated proliferation of conjunctival goblet cells in culture. Invest Ophthalmol Vis Sci 49:3351-359 CrossRef
18. Shatos MA, Rios JD, Horikawa Y, Hodges RR, Chang EL, Bernardino CR, Rubin PA, Dartt DA (2003) Isolation and characterization of cultured human conjunctival goblet cells. Invest Ophthalmol Vis Sci 44:2477-486 CrossRef
19. Chung SH, Lee JH, Yoon JH, Lee HK, Seo KY (2007) Multi-layered culture of primary human conjunctival epithelial cells producing MUC5AC. Exp Eye Res 85:226-33 CrossRef
20. Lee J, Jun EJ, Sunwoo JH, Kim ES, Kim JH, Kim JY, Kim MJ, Kim YK, Lee H, Tchah H (2011) An ex vivo model of coxsackievirus infection using multilayered human conjunctival epithelial cells. Graefes Arch Clin Exp Ophthalmol 249:1327-332 CrossRef
21. van Setten GB, Tervo T, Viinikka L, Perheentupa J, Tarkkanen A (1991) Epidermal growth factor in human tear fluid: a minireview. Int Ophthalmol 15:359-62 CrossRef
22. Enriquez-de-Salamanca A, Castellanos E, Stern ME, Fernández I, Carre?o E, García-Vázquez C, Herreras JM, Calonge M (2010) Tear cytokine and chemokine analysis and clinical correlations in evaporative-type dry eye disease. Mol Vis 16:862-73
23. Rao K, Farley WJ, Pflugfelder SC (2010) Association between high tear epidermal growth factor levels and corneal subepithelial fibrosis in dry eye conditions. Invest Ophthalmol Vis Sci 51:844-49 CrossRef
24. Luo L, Li DQ, Corrales RM, Pflugfelder SC (2005) Hyperosmolar saline is a proinflammatory stress on the mouse ocular surface. Eye Contact Lens 31:186-93 CrossRef
25. Corrales RM, Luo L, Chang EY, Pflugfelder SC (2008) Effects of osmoprotectants on hyperosmolar stress in cultured human corneal epithelial cells. Cornea 27:574-79 CrossRef
26. Li J, Zhu HY, Beuerman RW (2009) Stimulation of specific cytokines in human conjunctival epithelial cells by defensins HNP1, HBD2, and HBD3. Invest Ophthalmol Vis Sci 50:644-53 CrossRef
27. Adhikary G, Sun Y, Pearlman E (2008) C-Jun NH2 terminal kinase (JNK) is an essential mediator of Toll-like receptor 2-induced corneal inflammation. J Leukoc Biol 83:991-97 CrossRef
28. Yeh S, Song XJ, Farley W, Li DQ, Stern ME, Pflugfelder SC (2003) Apoptosis of ocular surface cells in experimentally induced dry eye. Invest Ophthalmol Vis Sci 44:124-29 CrossRef
29. Turner K, Pflugfelder SC, Ji Z, Feuer WJ, Stern M, Reis BL (2000) Interleukin-6 levels in the conjunctival epithelium of patients with dry eye disease treated with cyclosporine ophthalmic emulsion. Cornea 19:492-96 CrossRef
30. Luo L, Li DQ, Doshi A, Farley W, Corrales RM, Pflugfelder SC (2004) Experimental dry eye stimulates production of inflammatory cytokines and MMP-9 and activates MAPK signaling pathways on the ocular surface. Invest Ophthalmol Vis Sci 45:4293-301 CrossRef
31. Massingale ML, Li X, Vallabhajosyula M, Chen D, Wei Y, Asbell PA (2009) Analysis of inflammatory cytokines in the tears of dry eye patients. Cornea 28:1023-027 CrossRef
32. Singh AB, Harris RC (2005) Autocrine, paracrine and juxtacrine signaling by EGFR ligands. Cell Signal 17:1183-193 CrossRef
33. Grant MB, Khaw PT, Schultz GS, Adams JL, Shimizu RW (1992) Effects of epidermal growth factor, fibroblast growth factor, and transforming growth factor-beta on corneal cell chemotaxis. Invest Ophthalmol Vis Sci 33:3292-301
34. Fredj-Reygrobellet D, Plouet J, Delayre T, Baudouin C, Bourret F, Lapalus P (1987) Effects of aFGF and bFGF on wound healing in rabbit corneas. Curr Eye Res 6:1205-209 CrossRef
35. van Setten GB, Schultz GS, Macauley S (1994) Growth factors in human tear fluid and in lacrimal glands. Adv Exp Med Biol 350:315-19 CrossRef
36. Barton K, Nava A, Monroy DC, Pflugfelder SC (1998) Cytokines and tear function in ocular surface disease. Adv Exp Med Biol 438:461-69 CrossRef
37. Nezu E, Ohashi Y, Kinoshita S, Manabe R (1992) Recombinant human epidermal growth factor and corneal neovascularization. Jpn J Ophthalmol 36:401-06
38. Lu L, Wang L, Shell B (2003) UV-induced signaling pathways associated with corneal epithelial cell apoptosis. Invest Ophthalmol Vis Sci 44:5102-109 CrossRef
39. Krishnamoorthy M, Heimburg-Molinaro J, Bargo AM, Nash RJ (2009) Heparin binding epidermal growth factor-like growth factor and PD169316 prevent apoptosis in mouse embryonic stem cells. J Biochem 145:177-84 CrossRef
40. Li DQ, Tseng SC (1996) Differential regulation of cytokine and receptor transcript expression in human corneal and limbal fibroblasts by epidermal growth factor, transforming growth factor-alpha, platelet-derived growth factor B, and interleukin-1 beta. Invest Ophthalmol Vis Sci 37:2068-080 - 作者单位：Jae-hyung Kim (1)
Soon-Suk Kang (2)
Eun Soon Kim (2)
Jae Yong Kim (2)
Myoung Joon Kim (2)
Hungwon Tchah (2)
1. Department of Ophthalmology, Chungbuk National University College of Medicine, Cheongju, South Korea
2. Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, 388-1 Pungnap-2dong, Songpa-gu, Seoul, 138-736, South Korea
文摘
Background To analyze the effects of supplemental epidermal growth factor (EGF) and the roles of inflammatory cytokines (interleukin [IL]-6) in an ex vivo dry-eye model under hyperosmotic stress using a multilayered culture of human conjunctival epithelial cells (HCECs). Methods Multilayered cultures of HCECs were exposed to hyperosmotic stress (400?mOsm/L) for 24?h in addition to 0.5?ng/mL EGF (low-EGF group) or 25?ng/mL EGF (high-EGF group). Apoptosis was analyzed using the TUNEL assay. Cell proliferation was measured using the [3H]-thymidine incorporation assay. The expression of IL-6, EGF, EGF receptor (EGFR), and phosphorylated extracellular signal-regulated kinase (p-ERK) was measured by western blot analysis. The secretion of IL-6 was measured using ELISA. Western blot analysis was also performed using antibodies against cleaved caspase-3. Results The percentage of apoptotic cells was lower in the high-EGF group (6.7?%) than in the low-EGF group (10.3?%). The high-EGF group demonstrated increased proliferation (323.7 counts/min in the low-EGF group vs 649.1 counts/min in the high-EGF group). EGF induced higher phosphor-EGFR expression and upregulated p-ERK in HCECs. In addition, EGF significantly decreased the secretion of IL-6 and cleaved caspase-3 in HCECs. Conclusions The level of IL-6 was increased in the ex vivo HCEC dry-eye model that was under hyperosmotic stress. Supplemental EGF reduces the level of IL-6, decreases apoptosis, and increases proliferation. These findings indicate that EGF has potential as a therapeutic agent for the treatment of dry eyes.